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Semaphorin Profiling of Periodontal Fibroblasts and Osteoblasts

Louisiana State University Health Science Center, Department of Cell Biology and Anatomy, Center of Excellence in Oral and Craniofacial Biology, School of Dentistry, 1100 Florida Avenue, New Orleans, LA 70119, USA; tlalli{at}lsuhsc.edu

View larger version (53K): [in this window] [in a new window]   Figure 1. Periodontal cell segregation. Periodontal ligament (green) and gingival fibroblasts (red) were co-cultured and displayed little segregation after 24 hrs (A). In contrast, pre-osteoblasts (green) and gingival fibroblasts (red) displayed greater segregation when co-cultured for 24 hrs (B). The percentages of cell contacts made by cells with either similar or dissimilar cell types (for periodontal ligament and gingival fibroblasts and pre-osteoblasts) were quantified and compared for cells co-cultured for 2, 24, or 72 hrs (C). Data points represent the mean and standard deviation of 3 independent experiments, each consisting of 10 non-overlapping microscopic fields containing at least 50 cells each.

View larger version (45K): [in this window] [in a new window]   Figure 2. Expression of semaphorin, neuropilin, and plexin transcripts. (A) Reverse transcriptase polymerase chain-reaction (RT-PCR) analysis of transcripts expressed by gingival fibroblasts (G), dermal fibroblasts (D), periodontal ligament fibroblasts (P), and pre-osteoblasts (O). Relative RNA transcript levels for the semaphorins, neuropilins, and plexins were compared. The expression of the β1 subunit of integrin acts as a loading control, since it is expressed uniformly by all of the cells examined. The photo represents a typical result from 1 of 3 experiments. RT-PCR was performed for 25 cycles in this experiment. All RT-PCR reactions were repeated with 30 cycles for those transcripts that were initially detected poorly at 25 cycles. The PDL and gingival fibroblasts represented in panel (A) were isolated from the same individual. The expression of each transcript was compared between fibroblast populations (B) and fibroblast and pre-osteoblast populations (C). Four independent isolates of dermal, PDL, and gingival fibroblasts were compared, and the comparison of expression was represented as the ratio of the mean transcript expression. Error bars represent the standard deviation. In general, a five-fold difference in expression was determined to be statistically significant by ANOVA analysis. (D) Those semaphorins and plexins with a greater-than-five-fold difference in expression are listed.

View larger version (22K): [in this window] [in a new window]   Figure 3. Semaphorin and plexin protein expression in periodontal cells. The expression of Semaphorin 3A, Plexin A1, and Plexin C1 in dermal fibroblasts (DF), gingival fibroblasts (GF), periodontal ligament fibroblasts (PDLF), pre-osteoblasts (OB), and intact periodontal ligament (PDL) was examined by ELISA. Each bar represents the mean and standard deviation for 4 samples each for 4 cell/tissue isolates (n = 16).

Journal of Dental Research, Vol. 83, No. 9, 677-682 (2004)
DOI: 10.1177/154405910408300904


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